EP0155193A2 - Méthodes d'essai - Google Patents

Méthodes d'essai Download PDF

Info

Publication number
EP0155193A2
EP0155193A2 EP85301836A EP85301836A EP0155193A2 EP 0155193 A2 EP0155193 A2 EP 0155193A2 EP 85301836 A EP85301836 A EP 85301836A EP 85301836 A EP85301836 A EP 85301836A EP 0155193 A2 EP0155193 A2 EP 0155193A2
Authority
EP
European Patent Office
Prior art keywords
ligand
specific binding
binding partner
gate electrode
characteristic
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP85301836A
Other languages
German (de)
English (en)
Other versions
EP0155193A3 (en
EP0155193B1 (fr
Inventor
Gordon Coulter Forrest
Simon John Rattle
Grenville Arthur Robinson
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Merck Serono SA
Original Assignee
Serono Diagnostics Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Serono Diagnostics Ltd filed Critical Serono Diagnostics Ltd
Publication of EP0155193A2 publication Critical patent/EP0155193A2/fr
Publication of EP0155193A3 publication Critical patent/EP0155193A3/en
Application granted granted Critical
Publication of EP0155193B1 publication Critical patent/EP0155193B1/fr
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G01N33/5438—Electrodes
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/817—Enzyme or microbe electrode
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/806—Electrical property or magnetic property
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/807—Apparatus included in process claim, e.g. physical support structures

Definitions

  • the present invention relates to methods of assaying one of a pair of specific binding partners, and to apparatus for carrying out these methods.
  • the assay of ligand is determined by monitoring the extent of complex formation, for example by use of chemical or biochemical labels.
  • labelling employing radioisotopic, fluorescent or bioluminescent species, spin-labelling or enzyme labelling.
  • radioisotopic labels has become particularly widespread, due to the high degree of sensitivity and specificity obtainable.
  • an enzyme label which has been employed is an enzyme label.
  • the enzyme label is monitored for example by measuring the removal of substrate or the appearance of the product of the enzyme-catalysed substrate reaction by spectrophotometry, nephelometry, fluorimetry or by radiometry.
  • Such monitoring techniques may lack the high degree of sensitivity and specificity required for modern assay work. This may be due to the fact that neither the primary nor the secondary reaction is 100% quantitative or there may be inaccuracy in end-point assessment.
  • the use of radioactive labels presents the usual problems of safety and short shelf-life of the reagents.
  • the chromogens used in spectrophotometric techniques are often carcinogenic.
  • the apparatus used with known assay methods of the types mentioned above is not suitable for miniaturisation.
  • the methods thus require relatively large amounts of inconvenient or even dangerous reagents and the apparatus tends to be bulky and expensive.
  • FET field effect transistors
  • the invention provides a method of assaying a ligand in a sample, which comprises contacting the sample with a predetermined quantity of a specific binding partner to the ligand, said specific binding partner being immobilised on the effective gate electrode of a field effect transistor, and determining whether (and, if desired, the extent to which) an appropriate transistor characteristic is changed as a result of complex formation between the ligand and the specific binding partner.
  • the assay can be completed from the determined change in the transistor characteristic with reference to calibration data.
  • the method of the present invention is applicable both to qualitative and to quantitative assays.
  • effective gate electrode refers to an electrode functioning as the gate electrode of the FET. It will be appreciated that the construction of an effective gate electrode may differ considerably from that of conventional gate electrodes. The construction of effective gate electrodes is described below.
  • a predetermined quantity of a specific binding partner is immobilised on the effective gate electrode.
  • the specific binding partner may be present in a quantity greater than, equivalent to or less than the quantity of ligand present in the sample.
  • the change in the transistor characteristic may be determined as an absolute change observed after the complexing reaction has been allowed to proceed for a controlled period of time, or a rate of change of that characteristic with respect to time.
  • the initial rate of change may conveniently be measured. The method used will always, however, be consistent with the method employed in the calibration experiments.
  • the specific binding partner In the "absolute” mode, the specific binding partner will typically be present in a quantity greater than or equivalent to the quantity of ligand present in the sample.
  • the specific binding partner may be present in a quantity greater than, equivalent to or less than the quantity of ligand present in the sample.
  • a gate insulator 1 (preferably ion-insensitive) of insulating material [e.g. silicon dioxide, silicon nitride (which may for example have been treated with sodium hydroxide to reduce pH sensitivity) or paralene (which may for example have been deposited under vacuum thereby giving an ion-insensitive FET)] approximately 1000 A in thickness covers the surface of a p-type silicon substrate 2 between two diffusions of n-type silicon (the 'source' area S and 'drain' area D).
  • An effective gate electrode 3 on which is immobilised a predetermined quantity of the specific binding partner P lies over the gate insulator and an electrolyte 4 is present in contact with the electrode and the insulator.
  • V T threshold potential
  • the threshold potential VT of the FET will vary with individual devices. It can have a value of zero or can have a positive or negative non- zero value. As mentioned above, a suitable FET for use in the present invention may have a V T of approximately -0.5 V so that the device may be "on" without the immobilised specific binding partner being subjected to a large local electric field.
  • any change in the other characteristic will be due directly to complex formation between the specific binding partner and the ligand under assay. If in addition, the parameters of V d , pH, ionic strength and temperature are maintained constant and/or variations therein compensated for, such a change will be due solely to complex formation.
  • the pH may, for example, be controlled by the use of buffers.
  • a method of assay as herein described wherein the change in one transistor characteristic selected from the drain current characteristic and the gate electrode potential characteristic as a result of complex formation is determined, the other characteristic and optionally also the drain potential V d , the pH and ionic strength of the assay medium and the temperature of the system being maintained constant or variations therein compensated for.
  • the nature of the effective gate electrode may be varied to suit the assay conditions.
  • Particularly suitable for the assays of the present invention is a titanium dioxide gate electrode which may be chemically activated to permit coupling with the specific binding partner.
  • a conventional glass-bodied calomel electrode, a Ag/AgCl electrode, or any other suitable gate electrode may be used. Techniques for immobilising specific binding partners to glass and metal oxide surfaces are well-known in the art.
  • a reference electrode may act as a secondary gate electrode or may be used with a second FET, the outputs of the two transistors being monitored differentially. If appropriate, such an electrode may be chemically activated in an analagous way to the primary gate electrode, although not carrying specific binding partner. Furthermore, non-specific effects may be reduced by the covering the electrode bearing the immobilised specific binding partner with a membrane which is either permeable to, or perm-selective for, the ligand under assay. The reference system may also be protected by such a membrane.
  • apparatus for carrying out the assay methods as herein described, which apparatus includes a field effect transistor, the effective gate electrode of which has the specific binding partner immobilised thereon, and optionally a reference electrode.
  • An aqueous assay medium may be present, comprising a suitable pH buffer, which may advantageously act as an electrolyte. Means may be provided for incubating the assay mixture at any desired temperature.
  • the aqueous assay medium and the sample to be assayed may conveniently be introduced in any desired order through an entry port provided in the apparatus.
  • the apparatus will generally further comprise a suitable electrical circuit.
  • the apparatus may be pre-calibrated and provided with a scale whereby the change in the appropriate transistor characteristic may be read off directly as an amount of ligand in the sample.
  • the invention will be particularly described hereinafter with reference to an antibody or an antigen as a ligand.
  • the invention is not to be taken as being limited to assays of antibodies or antigens. Examples of ligands which may be assayed by the method of the invention are given in Table I below, together with an indication of a suitable specific binding partner in each instance.
  • the method of the invention is particularly suitable for assaying the following ligands: Hormones, including peptide hormones (e.g. thyroid stimulating hormone (TSH), lutenising hormone (LH), follicle stimulating hormone (FSH), insulin and prolactin) and non-peptide hormones (such as thyroxine and tri-iodothyronine), proteins, including human chorionic gonadotrophin (HCG), carcinoembryonic antigen (CEA ⁇ and alphafetoprotein (AFP), drugs (e.g. digoxin), sugars, toxins and vitamins.
  • TSH thyroid stimulating hormone
  • LH lutenising hormone
  • FSH follicle stimulating hormone
  • insulin and prolactin insulin and prolactin
  • non-peptide hormones such as thyroxine and tri-iodothyronine
  • proteins including human chorionic gonadotrophin (HCG), carcinoembryonic antigen (CEA ⁇ and alphafeto
  • antigen as used herein will be understood to include both permanently antigenic species (for example, proteins, bacteria, bacteria fragments, cells, cell fragments and viruses) and haptens which may be rendered antigenic under suitable conditions.
  • Immobilisation of an antibody or antigen molecule onto a gate electrode may be effected by various methods.
  • the attachment of the antibody or antigen to the electrode can be via any portion of the molecular structure so long as specific immunological activity is retained at the antibody or antigen binding site.
  • the electrode and/or the antibody or antigen may be chemically modified to permit successful attachment.
  • a titanium dioxide electrode carrying immobilised antibody may be prepared by heating a titanium wire at about 1000°C to form a thin oxide layer on its surface, contacting it firstly with a basic solution of cyanogen bromide and then with a basic solution of the antibody, and finally deactivating with urea the remaining active sites which have not taken up antibody.
  • a titanium dioxide electrode carrying immobilised protein e.g.
  • an enzyme such as glucose oxidase may be prepared by heating a titanium wire at about 900 - 1000 6 C and allowing it to cool to form a thin titanium dioxide layer on its surface, silanising it (for example by contacting with a solution of X-aminopropyltriethoxysilane in an appropriate solvent, e.g. acetone and subsequently evaporating the solvent) and then contacting it with firstly a solution of glutaraldehyde and secondly with a solution of the protein, and finally washing and deactivating the remaining active sites with urea.
  • a reference electrode may be prepared in analogous ways but omitting the stage of contacting the wire with the solution of the antibody or protein.
  • Electrode-immobilisation of an antibody or antigen reagent may also for example be achieved by bonding interactions between functional groups on the antibody or antigen molecule and the electrode, or by cross-linking or adsorption onto the surface of the electrode. Binding of reagents to the electrode may generally be accomplished by methods analogous to known methods of binding such reagents to solid supports (e.g. particles and beads), for example those described in European Patent Application No. 83305834.0.
  • the magnitude of the change in transistor characteristic due to complexing between an antibody and antigen will typically be of the order of millivolts or milliamperes.
  • the magnitude may vary due, inter alia, to the particular electrode used, the pH of the medium, the purity of the reagent used on the electrode and the ionic strength of the medium, and such parameters must therefore be consistent between the calibration experiments and the assay methods.
  • Titanium rods (2 cm long x 0.2 cm diameter) were heated for 12 hours at +900°C and then allowed - to cool overnight, thus growing a thin layer of titanium dioxide on the surface.
  • Silanisation of the rods was achieved by adding them to a 1% (v/v) solution of ⁇ -aminopropyltriethoxysilane in acetone and evaporating to dryness. The rods were then baked at 80°C for 22 hours.
  • the rods were soaked in a 2.5% (v/v) solution of- glutaraldehyde in phosphate buffer (50 mM, pH 7.4) for 22 hours at 25°C. After thorough washing with deionised water, the rods were contacted with a solution of glucose oxidase (0.33 mg/ml in phosphate buffer, 50 mM, pH 7.4)'overnight at 4°C. After thorough washing with deionised water and soaking with 6M urea to remove any adsorbed enzyme the rods were assayed for glucose oxidate activity.
  • a control rod was produced by following the above procedure but not contacting the rod with glucose oxidase.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
EP85301836A 1984-03-16 1985-03-15 Méthodes d'essai Expired - Lifetime EP0155193B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB848406955A GB8406955D0 (en) 1984-03-16 1984-03-16 Assay
GB8406955 1984-03-16

Publications (3)

Publication Number Publication Date
EP0155193A2 true EP0155193A2 (fr) 1985-09-18
EP0155193A3 EP0155193A3 (en) 1989-01-25
EP0155193B1 EP0155193B1 (fr) 1992-06-24

Family

ID=10558221

Family Applications (1)

Application Number Title Priority Date Filing Date
EP85301836A Expired - Lifetime EP0155193B1 (fr) 1984-03-16 1985-03-15 Méthodes d'essai

Country Status (8)

Country Link
US (1) US4778769A (fr)
EP (1) EP0155193B1 (fr)
AU (1) AU578075B2 (fr)
CA (1) CA1241695A (fr)
DE (1) DE3586254T2 (fr)
GB (1) GB8406955D0 (fr)
IL (1) IL74570A (fr)
ZA (1) ZA851971B (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2599844A1 (fr) * 1986-06-09 1987-12-11 Centre Nat Rech Scient Phase solide conductrice pour dosage immunoenzymatique, son procede de preparation et son utilisation dans des procedes de dosage immunoenzymatique
WO1988008875A1 (fr) * 1987-05-05 1988-11-17 The Washington Technology Center Detecteur a biopuce a action repetee
EP0245477A4 (fr) * 1985-11-19 1989-08-09 Univ Johns Hopkins Capteur capacitif d'analyse et de mesure chimiques.
US4927502A (en) * 1989-01-31 1990-05-22 Board Of Regents, The University Of Texas Methods and apparatus using galvanic immunoelectrodes
EP0328380A3 (en) * 1988-02-10 1990-06-27 Nec Corporation Electrochemical sensors in immunological measurement
EP0395137A3 (fr) * 1989-04-21 1991-06-12 ENIRICERCHE S.p.A. Capteur comprenant un antigène lié chimiquement à un dispositif à semi-conducteur
US5074977A (en) * 1987-05-05 1991-12-24 The Washington Technology Center Digital biosensors and method of using same
WO2013008062A1 (fr) * 2011-07-12 2013-01-17 Centre National De La Recherche Scientifique (Cnrs) Structure moléculaire non covalente, comprenant un glycoconjugué à base de pyrène, dispositif la comprenant et son utilisation pour la détection d'une lectine

Families Citing this family (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0315788B1 (fr) * 1987-10-13 1992-12-09 Taiyo Yuden Co., Ltd. Senseur à ions
US4973910A (en) * 1988-01-14 1990-11-27 Wilson Mahlon S Surface potential analyzer
US5057430A (en) * 1988-09-15 1991-10-15 Biotronic Systems Corporation Biochemical sensor responsive to bubbles
US5200051A (en) * 1988-11-14 1993-04-06 I-Stat Corporation Wholly microfabricated biosensors and process for the manufacture and use thereof
US6306594B1 (en) 1988-11-14 2001-10-23 I-Stat Corporation Methods for microdispensing patterened layers
AU9113991A (en) * 1990-11-19 1992-06-11 Biotechnology Research And Development Corporation Mutant orientable proteins and coated substrates
US5494831A (en) * 1993-08-30 1996-02-27 Hughes Aircraft Company Electrochemical immunosensor system and methods
US5641634A (en) * 1995-11-30 1997-06-24 Mandecki; Wlodek Electronically-indexed solid-phase assay for biomolecules
US6051377A (en) * 1995-11-30 2000-04-18 Pharmaseq, Inc. Multiplex assay for nucleic acids employing transponders
AU1061997A (en) 1995-11-30 1997-06-19 Wlodek Mandecki Screening of drugs from chemical combinatorial libraries employing transponders
US6001571A (en) * 1995-11-30 1999-12-14 Mandecki; Wlodek Multiplex assay for nucleic acids employing transponders
US5736332A (en) * 1995-11-30 1998-04-07 Mandecki; Wlodek Method of determining the sequence of nucleic acids employing solid-phase particles carrying transponders
US5981166A (en) * 1997-04-23 1999-11-09 Pharmaseq, Inc. Screening of soluble chemical compounds for their pharmacological properties utilizing transponders
JP3612675B2 (ja) 1997-06-05 2005-01-19 日本光電工業株式会社 ｐＨ変化誘因性物質または生物体の測定装置及び測定方法
US5940212A (en) * 1997-10-30 1999-08-17 3M Innovative Properties Company Cube corner article with altered inactive areas and method of making same
EP1030912A4 (fr) 1997-11-14 2007-01-10 California Inst Of Techn Dispositif servant a effectuer la lyse de cellules
AU2002311951A1 (en) * 2001-05-18 2002-12-03 Regents Of The University Of Minnesota Metal/metal oxide electrode as ph-sensor and methods of production
US6653653B2 (en) 2001-07-13 2003-11-25 Quantum Logic Devices, Inc. Single-electron transistors and fabrication methods in which a projecting feature defines spacing between electrodes
US6483125B1 (en) 2001-07-13 2002-11-19 North Carolina State University Single electron transistors in which the thickness of an insulating layer defines spacing between electrodes
DE10163557B4 (de) * 2001-12-21 2007-12-06 Forschungszentrum Jülich GmbH Transistorbasierter Sensor mit besonders ausgestalteter Gateelektrode zur hochempfindlichen Detektion von Analyten
US6673717B1 (en) 2002-06-26 2004-01-06 Quantum Logic Devices, Inc. Methods for fabricating nanopores for single-electron devices
GB0322010D0 (en) * 2003-09-19 2003-10-22 Univ Cambridge Tech Detection of molecular interactions using field effect transistors
JP3874772B2 (ja) * 2004-07-21 2007-01-31 株式会社日立製作所 生体関連物質測定装置及び測定方法
JP4081477B2 (ja) * 2005-03-29 2008-04-23 株式会社日立製作所 生体分子検出装置及びそれを用いた生体分子検出方法
US8653839B2 (en) * 2009-07-17 2014-02-18 Caterpillar Inc. Zinc oxide sulfur sensors and method of using said sensors
US8373206B2 (en) 2010-07-20 2013-02-12 Nth Tech Corporation Biosensor apparatuses and methods thereof
US10378044B1 (en) * 2013-10-01 2019-08-13 FemtoDx Methods for increasing the molecular specificity of a nanosensor

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ZA704115B (en) * 1969-06-23 1971-03-31 Miles Lab Apparatus and method for the determination of substances in test reaction systems
US4020830A (en) * 1975-03-12 1977-05-03 The University Of Utah Selective chemical sensitive FET transducers
SE407115B (sv) * 1975-10-06 1979-03-12 Kabi Ab Forfarande och metelektroder for studium av enzymatiska och andra biokemiska reaktioner
US4238757A (en) * 1976-03-19 1980-12-09 General Electric Company Field effect transistor for detection of biological reactions
US4273636A (en) * 1977-05-26 1981-06-16 Kiyoo Shimada Selective chemical sensitive field effect transistor transducers
JPS5539042A (en) * 1978-09-14 1980-03-18 Tokyo Daigaku Ion selecting field-effect sensor
GB2096824A (en) * 1981-04-09 1982-10-20 Sibbald Alastair Chemically sensitive field effect transistor
US4411741A (en) * 1982-01-12 1983-10-25 University Of Utah Apparatus and method for measuring the concentration of components in fluids
SE8200442L (sv) * 1982-01-27 1983-07-28 Forsvarets Forsknings Forfarande vid detektion av organiska molekyler, sasom biomolekyler
GB8314523D0 (en) * 1983-05-25 1983-06-29 Lowe C R Diagnostic device
AU596946B2 (en) * 1984-07-10 1990-05-24 F. Hoffmann-La Roche Ltd Methods of assay

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0245477A4 (fr) * 1985-11-19 1989-08-09 Univ Johns Hopkins Capteur capacitif d'analyse et de mesure chimiques.
FR2599844A1 (fr) * 1986-06-09 1987-12-11 Centre Nat Rech Scient Phase solide conductrice pour dosage immunoenzymatique, son procede de preparation et son utilisation dans des procedes de dosage immunoenzymatique
WO1987007725A1 (fr) * 1986-06-09 1987-12-17 Centre National De La Recherche Scientifique (Cnrs Procede cyclique de dosage immunoenzymatique d'un ligand
WO1988008875A1 (fr) * 1987-05-05 1988-11-17 The Washington Technology Center Detecteur a biopuce a action repetee
US5074977A (en) * 1987-05-05 1991-12-24 The Washington Technology Center Digital biosensors and method of using same
EP0328380A3 (en) * 1988-02-10 1990-06-27 Nec Corporation Electrochemical sensors in immunological measurement
US4927502A (en) * 1989-01-31 1990-05-22 Board Of Regents, The University Of Texas Methods and apparatus using galvanic immunoelectrodes
EP0395137A3 (fr) * 1989-04-21 1991-06-12 ENIRICERCHE S.p.A. Capteur comprenant un antigène lié chimiquement à un dispositif à semi-conducteur
US5160597A (en) * 1989-04-21 1992-11-03 Eniricerche S.P.A. Sensor with antigen chemically bonded to a semiconductor device
WO2013008062A1 (fr) * 2011-07-12 2013-01-17 Centre National De La Recherche Scientifique (Cnrs) Structure moléculaire non covalente, comprenant un glycoconjugué à base de pyrène, dispositif la comprenant et son utilisation pour la détection d'une lectine

Also Published As

Publication number Publication date
ZA851971B (en) 1986-05-28
US4778769A (en) 1988-10-18
DE3586254T2 (de) 1993-02-11
AU4000285A (en) 1985-09-19
IL74570A (en) 1990-02-09
CA1241695A (fr) 1988-09-06
IL74570A0 (en) 1985-06-30
GB8406955D0 (en) 1984-04-18
DE3586254D1 (de) 1992-07-30
EP0155193A3 (en) 1989-01-25
EP0155193B1 (fr) 1992-06-24
AU578075B2 (en) 1988-10-13

Similar Documents

Publication Publication Date Title
US4778769A (en) Methods of electrochemical assay employing a field effect transistor
EP0163312A2 (fr) Immunoessai
Meyerhoff et al. Novel nonseparation sandwich-type electrochemical enzyme immunoassay system for detecting marker proteins in undiluted blood
EP0207152B1 (fr) Analyse de diffusion dans une phase solide
JPH0239747B2 (fr)
US4670383A (en) Immune-chemical measurement process for haptens and proteins
EP0149602A1 (fr) Analyse immunometrique utilisant des anticorps polyclonaux et monoclonaux et kit d'analyse
Robinson et al. Bioelectrochemical immunoassay for human chorionic gonadotrophin in serum using an electrode-immobilised capture antibody
KR100231628B1 (ko) 당단백질의 분리 및 측정 방법
US5236849A (en) Method of high sensitivity immunoassay
EP0303229B1 (fr) Essai immunologique très sensible
Boitieux et al. Oxygen electrode-based enzyme immunoassay for the amperometric determination of hepatitis B surface antigen
JPH081438B2 (ja) 酵素免疫測定法
IE850007L (en) Process and reagent for the determination of a polyvalent¹antigen
JPH0285755A (ja) 免疫センサ
EP0155135B1 (fr) Méthodes d'essai
EP0125368B1 (fr) Méthode de détection immunologique par potentiométrie de la vitesse de réaction en utilisant une électrode à sélectivité ionique au fluorure
EP0488195B1 (fr) Essai immunologique chemiluminescente
Anderson et al. Internally referenced ImmunoConcentration assays.
JP2000180446A (ja) 干渉作用を回避する方法及び試薬
Karube et al. A catalytic immuno-reactor for the amperometric determination of human serum albumin
JPH052940B2 (fr)
JPH08166388A (ja) 酵素免疫測定方法及び酵素免疫センサ
EP0345777B1 (fr) Immunoessai
JP2816767B2 (ja) 固定化抗体の製造方法

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Designated state(s): BE CH DE FR GB IT LI NL SE

PUAL Search report despatched

Free format text: ORIGINAL CODE: 0009013

AK Designated contracting states

Kind code of ref document: A3

Designated state(s): BE CH DE FR GB IT LI NL SE

17P Request for examination filed

Effective date: 19890717

17Q First examination report despatched

Effective date: 19901123

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): BE CH DE FR GB IT LI NL SE

ITF It: translation for a ep patent filed
REF Corresponds to:

Ref document number: 3586254

Country of ref document: DE

Date of ref document: 19920730

ET Fr: translation filed
PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed
ITTA It: last paid annual fee
EAL Se: european patent in force in sweden

Ref document number: 85301836.4

REG Reference to a national code

Ref country code: GB

Ref legal event code: 732E

REG Reference to a national code

Ref country code: CH

Ref legal event code: PUE

Owner name: SERONO DIAGNOSTICS LIMITED TRANSFER- SERONO DIAGNO

NLS Nl: assignments of ep-patents

Owner name: LABORATOIRES SERONO S.A.;SERONO DIAGNOSTIC S.A.

REG Reference to a national code

Ref country code: FR

Ref legal event code: TP

Ref country code: FR

Ref legal event code: CD

BECA Be: change of holder's address

Free format text: 960508 S.A. LABORATOIRES *SERONO:ZONE INDUSTRIELLE DE L'OURIETTAZ, 1170 AUBONNE

BECH Be: change of holder

Free format text: 960508 S.A. LABORATOIRES *SERONO

REG Reference to a national code

Ref country code: GB

Ref legal event code: IF02

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 20040304

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20040309

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20040310

Year of fee payment: 20

Ref country code: GB

Payment date: 20040310

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 20040317

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20040325

Year of fee payment: 20

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 20040518

Year of fee payment: 20

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LI

Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION

Effective date: 20050314

Ref country code: GB

Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION

Effective date: 20050314

Ref country code: CH

Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION

Effective date: 20050314

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Free format text: LAPSE BECAUSE OF EXPIRATION OF PROTECTION

Effective date: 20050315

BE20 Be: patent expired

Owner name: S.A. LABORATOIRES *SERONO

Effective date: 20050315

REG Reference to a national code

Ref country code: GB

Ref legal event code: PE20

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

NLV7 Nl: ceased due to reaching the maximum lifetime of a patent

Effective date: 20050315

EUG Se: european patent has lapsed
BE20 Be: patent expired

Owner name: S.A. LABORATOIRES *SERONO

Effective date: 20050315